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© β–Lactamase positive bacteria represent a growing threat to human health because of their resistance to commonly used antibiotics. Therefore, development of new diagnostic methods for identification of β–lactamase positive bacteria is of high importance for monitoring the spread of antibiotic–resistant bacteria. Here, we report the discovery of a new biodegradation metabolite (H2S), generated through β–lactamase–catalyzed hydrolysis of β–lactam antibiotics. This discovery directed us to develop a distinct molecular technique for monitoring bacterial antibiotic resistance. The technique is based on a highly efficient chemiluminescence probe, designed for detection of the metabolite, hydrogen sulfide, that is released upon biodegradation of β–lactam by β–lactamases. Such an assay can directly indicate if antibiotic bacterial resistance exists for a certain examined β–lactam. The assay was successfully demonstrated for five different β–lactam antibiotics and eight β–lactam resistant bacterial strains. Importantly, in a functional bacterial assay, our chemiluminescence probe was able to clearly distinguish between a β–lactam resistant bacterial strain and a sensitive one. As far as we know, there is no previous documentation for such a biodegradation pathway of β–lactam antibiotics. Bearing in mind the data obtained in this study, we propose that hydrogen sulfide should be considered as an emerging β–lactam metabolite for detection of bacterial resistance.
